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Identification of F. oxysporum sp. tuberosi as responsible of the corm rot of saffron and expression in field collected infected plants. A. Sequence comparison among the b-tubulin fragments from F. oxysporum f. sp. gladioli (Fo-gladioli) F. oxysporum f. sp. tuberosi (Fo-tuberosi) and the isolated fungi in this study (Fo). B. Saffron corms collected from outfields at different stages of damage, termed as number 1: healthy corm, 2: corm at an early stage of infection, 3: corm fully infected, and analysis of SafchiA expression in corms infected with Fusarium oxysporum f. sp. tuberosi at the same levels of pathogen proliferation as shown in the picture above. RT-PCR was done using a labelled fungal b-tubulin fragment as a probe to show fungal presence.
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In plants, various chitinases have been identified and categorized into several groups based on the analysis of their sequences and domains. We have isolated SafchiA, a novel class of chitinase from saffron (Crocus sativus L.). The cDNA encoding SafchiA is mainly expressed in roots and corms, and its expression is induced by elicitor treatment, met...
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Citations
... It was indicated that overexpression of the chitinase could enhance protection against fungal infection in transgenic tobacco 57 . López and Gómez-Gómez 58 demonstrated the promotion of the transcription of some chitinase genes under various abiotic stresses, including drought, cold, and salt 58 . ...
Biotic stress imposed by pathogens, including fungal, bacterial, and viral, can cause heavy damage leading to yield reduction in maize. Therefore, the identification of resistant genes paves the way to the development of disease-resistant cultivars and is essential for reliable production in maize. Identifying different gene expression patterns can deepen our perception of maize resistance to disease. This study includes machine learning and deep learning-based application for classifying genes expressed under normal and biotic stress in maize. Machine learning algorithms used are Naive Bayes (NB), K-Nearest Neighbor (KNN), Ensemble, Support Vector Machine (SVM), and Decision Tree (DT). A Bidirectional Long Short Term Memory (BiLSTM) based network with Recurrent Neural Network (RNN) architecture is proposed for gene classification with deep learning. To increase the performance of these algorithms, feature selection is made from the raw gene features through the Relief feature selection algorithm. The obtained finding indicated the efficacy of BiLSTM over other machine learning algorithms. Some top genes ((S)-beta-macrocarpene synthase, zealexin A1 synthase, polyphenol oxidase I, chloroplastic, pathogenesis-related protein 10, CHY1, chitinase chem 5, barwin, and uncharacterized LOC100273479 were proved to be differentially upregulated under biotic stress condition.
... Phoma species are reported from plants as pathogens (Rai and Misra 1981;Agrawal and Mishra 1981;Rao and Thirumalachar 1981;Davari and Hajieghrari 2008;Aveskamp et al. 2008;Lopez and Gomez-Gomez 2009;Pellegrino et al. 2010;Garibaldi et al. 2010;Quiroz et al. 2014;Gilardi et al. 2017;Colmán et al. 2018;Basavand et al. 2020; Yarmalovich and Siaredzich (Rai et al. 2018(Rai et al. , 2020, and soil (Mathur and Thirumalachar 1959;Dorenbosch 1970). There are few cases of Phoma infection in human beings (Shukla et al. 1984;Rai 1989Rai , 2004Suh 2005;Balis et al. 2006;Tullio et al. 2010;Kukhar et al. 2020). ...
Fabrication of metal nanoparticles till date has been reported by the use of several biological agents like bacteria, fungi, yeast, algae, and plants. Among all the biological systems used for the synthesis of metal nanoparticles, fungal system scores over the other biological system. The possible use of fungi has gained much importance, as they are easy to culture in bulk. Also, the extracellular secretion of biomolecules has an added advantage in the downstream processing and handling of biomass as well as reusability of biomass. The genus Phoma is a cosmopolitan fungus with a worldwide distribution. Various species of the genus Phoma produce numerous important secondary metabolites which include phytotoxins, antitumor, and anti-HIV agents.
... Moreover, chitinase-A from Cycas revoluta expresses additional transglycosylation activity which is unique in the plant kingdom [15]. The saffron chitinase from Crocus sativus [16] promotes the jasmonic acid signal pathway as defense activation mechanism. ...
Background
Chitin is one of the most abundant biopolymers on Earth, only trailing second after cellulose. The enzyme chitinase is responsible for the degradation of chitin. Chitinases are found to be produced by wide range of organisms ranging from archaea to higher plants. Though chitin is a major component of fungal cell walls and invertebrate exoskeletons, bacterial chitinase can be industrially generated at low cost, in facile downstream processes at high production rate. Microbial chitinases are more stable, active, and economically practicable compared to the plant- and animal-derived enzymes.
Results
In the present study, computationally obtained results showed functional characteristics of chitinase with particular emphasis on bacterial chitinase which is fulfilling all the required qualities needed for commercial production. Sixty-two chitinase sequences from four different groups of organisms were collected from the RCSB Protein Data Bank. Considering one suitable exemplary sequence from each group is being compared with others. Primary, secondary, and tertiary structures are determined by in silico models. Different physical parameters, viz., pI, molecular weight, instability index, aliphatic index, GRAVY, and presence of functional motifs, are determined, and a phylogenetic tree has been constructed to elucidate relationships with other groups of organisms.
Conclusions
This study provides novel insights into distribution of chitinase among four groups and their characterization. The results represent valuable information toward bacterial chitinase in terms of the catalytic properties and structural features, can be exploited to produce a range of chitin-derived products.
... [62] are also reported to inhibit growth of fungus in cocoa leaf, potato cyst and potato seed tubers, respectively. Similar inhibitory effects of chitinase obtained from plant sources have also been previously reported against different fungal species [21,45,63,64]. ...
Background
Chitin is a unique structural exopolysaccharide abundantly found in nature. This exopolysaccharide has a unique chemical structure that acts as a protective outermost covering for most of the crustaceans in aquatic ecosystem. This fortification is because of the insoluble nature of this exopolysaccharide which consist of a linear chain of β-(1→4)-linked-N-acetylglucosamine units. Chitin is hydrolyzed with the help of a hydrolase known as chitinase. Variety of microbial species have been explored for chitinase production. Chitinolytic microbial species can be alternatively used for degradation of chitin instead of chemical treatment in agricultural sector. This biological approach has lesser environmental impact because of its apparently safe nature.
Result
In the current study, bioprospecting of chitinase producing species was conducted and different chitinolytic bacterial strains were screened for chitinase production which could have anti-fungal potential. Bacterial isolates were identified based on polyphasic approach and the enzyme production was optimized using one-variable-at-a-time technique. Hyphal extension method was used for determination of anti-fungal potential of chitinase.Glutamicibacter uratoxydans was indigenously isolated and identified for chitinase production. G. uratoxydans is a novel bacterial species which has not been previously explored to produce chitinase or other hydrolases. G. uratoxydans biosynthesized chitinase utilizing colloidal chitin as a sole source of carbon. The chitinase biosynthesized by G. uratoxydans is effectively potent against Aspergillus fumigatus thus, suggesting that this extracellular enzyme could be used for the treatment of fungal infection caused by filamentous fungi.
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... Plant LOX may involve in growth and developmental control processes, through biosynthesis of regulatory molecules and volatile compounds involved in insect attraction, defense responses to pathogen, wounding and stresses [8]. cDNA encoding chitinase is mainly expressed in infected roots and corms, suggesting a defense role of protein to develop a range of defense-related enzymes, capable of initiating resistance to pathogens [31,68]. Wang et al. [61], performed molecular characterization and expression analysis of WRKY family genes in Dendrobium officinale. ...
... The corms were placed on pots and in the fields of Jardín Botánico de Castilla-la Mancha (Albacete, Spain). Leaves, roots, corms, and flowers were collected at the developmental stages previously described (Rubio et al., 2008;Lopez and Gomez-Gomez, 2009), frozen in liquid nitrogen and stored at −80 • C until further use. ...
Crocus sativus stigmas are the main source of crocins, which are glucosylated apocarotenoids derived from zeaxanthin cleavage that give saffron its red color. Phytoene synthase (PSY) mediates the first committed step in carotenoid biosynthesis in plants. Four PSY genes encoding functional enzymes were isolated from saffron. All the proteins were localized in plastids, but the expression patterns of each gene, CsPSY1a, CsPSY1b, CsPSY2, and CsPSY3, in different saffron tissues and during the development of the stigma showed different tissue specialization. The CsPSY2 transcript was primarily detected in the stigmas where it activates and stimulates the accumulation of crocins, while its expression was very low in other tissues. In contrast, CsPSY1a and CsPSY1b were mainly expressed in the leaves, but only CsPSY1b showed stress-light regulation. Interestingly, CsPSY1b showed differential expression of two alternative splice variants, which differ in the intron retention at their 5′ UTRs, resulting in a reduction in their expression levels. In addition, the CsPSY1a and CsPSY1b transcripts, together with the CsPSY3 transcript, were induced in roots under different stress conditions. The CsPSY3 expression was high in the root tip, and its expression was associated with mycorrhizal colonization and strigolactone production. CsPSY3 formed a separate branch to the stress-specific Poaceae homologs but was closely related to the dicot PSY3 enzymes.
... In addition, some plant hormones and abiotic stress also regulate the expression and activity of chitinases [27][28][29]. For instances, jasmonates and wounding can induce the expression of BjCHI and SafChia genes [30,31]. Some chitinases are upregulated by heavy metals including cadmium, and arsenic [32]. ...
... Some chitinases are upregulated by heavy metals including cadmium, and arsenic [32]. Moreover, salt, cold and drought stresses promote the transcription of some chitinase genes [28,31]. All these suggest that chitinases may be involved in a variety of growth processes and stress responses. ...
Chitinase catalyzes the hydrolysis of chitin β-1,4 linkages. However, plants cannot produce chitin, suggesting that plant chitinases do not have the same function as animals. This study investigated the chitinase gene family in tomato and divided into eight groups via phylogenetic analyses with Arabidopsis and rice members. Conserved gene structures and motif arrangements indicated their functional relevance with each group. These genes were nonrandomly distributed across the tomato chromosomes, and tandem duplication contributed to the expansion of this gene family. Synteny analysis also established orthology relationships and functional linkages between Arabidopsis and tomato chitinase genes. Several positive selection sites were identified, which may contribute to the functional divergence of the protein family in evolution. In addition, differential expression profiles of the tomato chitinase genes were also investigated at some developmental stages, or under different biotic and abiotic stresses. Finally, functional network analysis found 124 physical or functional interactions, implying the diversity of physiological functions of the family proteins. These results provide a foundation for the exploration of the chitinase genes in plants and will offer some insights for further functional studies.
... Crocus sativus (López & Gómez, 2009), from latex of Ipomoa carnea (Patel et al., 2010), Morus alba (Kitajima et al., 2010), Ficus microcarpa ( (Taira, Ohdomari et al., 2005), Carica papaya, Ficus carica, Euphorbia characias (Span et al., 2015) and Hevea brasiliensis (Martin, 1991), from leaves of tobacco, cotton, tea (Nisha, Prabu, Manda, & Arvinth, 2011), sugar beet, alfa alfa (Zhong & Hiruki, 1993), pokeweed (Ohta, Yamagami, & Funatsu, 1995), potato, Pteris ryukyuensis, beans and (Sorensen et al., 2010), from fruits of Diospyros kaki (Zhang, Kopparapu, Yan, Yang, & Jiang, 2013), Punica granatum , Ananas comosus (Taira, Ohdomari et al., 2005), ...
Easier susceptibility of plants toward vulnerable pathogens due to lack of an immune system necessitated evolution of different kind of responses priming the release of pathogenesis‐related (PR) defense proteins. With reference to attack of fungal pathogens, the lytic enzyme known as chitinase (EC 3.2.1.14) could easily degrade the fungal cell wall chitin and played a crucial role in defending plants. In the present review, various chitinases purified from different plant sources with elaboration of their properties have been explored in detail. Purification studies of chitinases showed a diverse range of molecular mass (25–40 kDa), optimum pH (4.0–6.0), temperature (50–60°C), and kinetic parameters. Along with this, purified chitinases also showed strong antifungal activity against phytopathogenic fungi and nonpathogenic plant fungi.
Practical applications
The present review provides a single platform for all the purification protocols adopted for purification of plant chitinases in a single article. It results into simplification of study of plant chitinases purified from different parts of plants so far. For the readers, who want to be benefitted with the summarized study of plant chitinases and their properties, this review article fulfills their need and quite helpful in context of other review article. Along with this, the review emphasizes on the antifungal properties of plant‐purified chitinases and seems to be helpful for those who want to employ these chitinases as a biocontrol agent at larger scale in managing plant fungal diseases.
... However, it was devoid of antifungal activity against Alternaria alternata even at concentration of 4.20 μg/ml (Figure 5c). Higher plants contain both constitutive and inducible defense systems (Lopez & Gomez, 2009). Usually, a constitutive response is stable and nonspecific. ...
A chitinase was purified from naked oat (Avena chinensis) seeds using simple chromatographic techniques. Its molecular weight and isoelectric point were determined as 35 kDa and 8.9, respectively. The purified chitinase exhibited specific activity of 3.6 U/mg and 15.6% yield using colloidal chitin as substrate. Partial amino acid sequence analysis and homology search indicated that it probably belonged to Class I plant chitinase, glycosyl hydrolase family 19. With chitin as substrate, the optimum pH and temperature of the chitinase were pH 7.0 and 40°C, respectively. The chitinase was remarkably stable from 30°C up to 50°C, but was inactivated at high temperatures above 85°C. Antifungal activity in vitro tests demonstrated this purified chitinase had potent, dose‐dependent inhibitory activity against the fungi Panus conchatus and Trichoderma reesei.
Practical applications
Chitinase has broad applications in many fields including the food industry and is recognized as one of the antifungal substances with potential use in plant disease resistance or biological control in agriculture. This study developed cost‐effective purification methods for producing chitinase from naked oat (Avena chinensis) seeds, which may favor large‐scale production of the enzyme. The remarkable stability of the chitinase at moderate temperatures (30°C–50°C), makes it a potentially useful enzyme in bioprocessing to produce chitooligosaccharides for various applications in the food, health, and agriculture sectors.
... Iridaceae comprised of traditional medicinal plants as rich source of secondary metabolites [12]. In natural circumstances, the seeds and corms are frequently attacked by various pathogens including viruses, bacteria, fungi and nematodes [13]. Various chitinase enzymes have received attention due to their broad applications in biotechnology, medicine, waste management and other industries as well as in agriculture for biocontrol of phytopathogenic fungi and harmful insects [14]. ...
The present study describes the predicted model and functional characterization of
an endochitinase (30 kDa) from corms of Gladiolus grandiflorus. ESI-QTOF-MS generated peptide showed 96% sequence homology with family 18, Class III acidic endochitinase of Gladiolus
gandavensis. Purified G. grandiflorus chitinase (GgChi) hydrolyzed 4-methylumbelliferyl b-DN,N0,N00-triacetylchitotriose substrate showing specific endochitinase activity. Since no structural details of GgChi were available in the Protein Data Bank (PDB), a homology model was
predicted using the coordinate information of Crocus vernus chitinase (PDB ID: 3SIM). Ramachandran plot indicated 84.5% in most favored region, 14.8% in additional and 0.6% in generously allowed region while no residue in disallowed region. The predicted structure indicated a highly conserved (b/a)8 (TIM barrel) structure similar to the family 18, class III chitinases. The GgChi also showed sequence and structural homologies with other active
chitinases. The GgChi (50 mg/disc) showed no antibacterial activity, but did provide mild
growth inhibition of phytopathogenic fungus Fusarium oxysporum at a concentration of 500
mg/well Similarly, insect toxicity bioassays of GgChi (50 mg) against nymphs of Bemisia tabaci
showed 14% reduction in adult emergence and 14% increase in mortality rate in comparison to
control values. The GgChi (1.5 mg) protein showed significant reduction in a population of flour
beetle (Tribolium castaneum) after 35 days, but lower reactivity against rice weevil (Sitophilus
oryzae). The results of this study provide detai.led insight on functional characterization of a
family 18 class III acidic plant endochitinase.
